过表达MyBPC1对牛骨骼肌卫星细胞增殖分化的影响

doi:10.11843/j.issn.0366-6964.2021.012.011

摘要/Abstract

摘要： 旨在探究肌球蛋白结合蛋白C1（myosin binding protein C1，MyBPC1）对牛骨骼肌卫星细胞增殖与成肌分化的影响，为进一步研究MyBPC1在细胞分化和肌肉发育过程中的调控作用提供依据。本研究利用西门塔尔胎牛原代牛骨骼肌卫星细胞体外诱导成肌分化模型模拟牛骨骼肌的生长发育过程。采用qRT-PCR和Western blot检测MyBPC1的细胞时序表达谱。试验分为两组。在RNA水平每组4个重复，每个重复20 μL；在蛋白水平每组3个重复，每个重复15 μg。采用qRT-PCR和Western blot检测牛骨骼肌卫星细胞转染MyBPC1的过表达效果，并进一步检测细胞增殖期标志因子Pax7、Ki67以及细胞分化期标志因子MyHC、MyOG的表达变化情况，观察牛骨骼肌卫星细胞肌管形成状态。结果，MyBPC1在牛骨骼肌卫星细胞分化前后表达水平存在极显著差异，牛骨骼肌卫星细胞诱导分化后MyBPC1的mRNA和蛋白表达量均极显著高于增殖期（P<0.01）。过表达MyBPC1后，细胞分化形成的肌管数量明显多于对照组，增殖标志因子Pax7的mRNA水平和蛋白表达水平无显著差异，分化标志因子MyHC的mRNA水平和蛋白表达水平极显著高于对照组（P<0.01）。过表达MyBPC1可以促进牛骨骼肌卫星细胞体外成肌分化，为进一步开展MyBPC1对牛骨骼肌卫星细胞的调控机制奠定基础。

关键词: 牛, 骨骼肌卫星细胞, 过表达, MyBPC1, 增殖, 分化

Abstract: The study aimed to explore the effects of myosin binding protein C1 (MyBPC1) on the proliferation and myogenic differentiation of bovine skeletal muscle satellite cells, and provide a basis for further study of the regulatory role of MyBPC1 in cell differentiation and muscle development. The Simmental fetal bovine primary bovine skeletal muscle satellite cells to inducing myogenic differentiation model in vitro was used to simulate the growth and development process of bovine skeletal muscle. qRT-PCR and Western blot were used to detect the cellular temporal expression profile of MyBPC1. There were two groups in this experiment. At the RNA level, there were 4 replicates in each group, and 20 μL in each replicate. However, at the protein level, there were 3 replicates in each group, and 15 μg in each replicate. qRT-PCR and Western blot were used to detect the overexpression effect of MyBPC1 transfected into bovine skeletal muscle sate-llite cells, and to further detect the expression changes of cell proliferation marker factors Pax7, Ki67 and cell differentiation marker factors MyHC, MyOG. The myotube formation status of bovine skeletal muscle satellite cells was observed. The expression level of MyBPC1 was significantly different before and after the differentiation of bovine skeletal muscle satellite cells. After the differentiation of bovine skeletal muscle satellite cells, the mRNA and protein expression of MyBPC1 were significantly higher than those in the proliferation phase (P<0.01). After overexpression of MyBPC1, the number of myotubes formed by cell differentiation was significantly greater than that of the control group. There was no significant difference at the mRNA and protein expression levels of the proliferation marker factor Pax7. The mRNA and protein expression levels of the differentiation marker factor MyHC were significantly higher than that of the control group (P<0.01). Overexpression of MyBPC1 can promote the myogenic differentiation of bovine skele-tal muscle satellite cells in vitro, which lays the foundation for further studying the regulatory mechanism of MyBPC1 on bovine skeletal muscle satellite cells.

Key words: bovine, skeletal muscle satellite cells, overexpression, MyBPC1, proliferation, differentiation

中图分类号:

S823.2

洪倩倩, 郭宏, 高树新, 郭益文. 过表达MyBPC1对牛骨骼肌卫星细胞增殖分化的影响[J]. 畜牧兽医学报, 2021, 52(12): 3439-3448.

HONG Qianqian, GUO Hong, GAO Shuxin, GUO Yiwen. Effect of Overexpression of MyBPC1 on Proliferation and Differentiation of Bovine Skeletal Muscle Satellite Cells[J]. Acta Veterinaria et Zootechnica Sinica, 2021, 52(12): 3439-3448.

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